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MERC+GSI – DRAFT Test Plan
8
between the two sets of tests. The following sections describe how each parameter and variable
is sampled/analyzed and additional details can be found in Appendices and at
www.nemw.org/GSI/SOPS.htm.
5. Methods
Quantifying Physical Conditions:
Temperature, salinity, dissolved oxygen, chlorophyll fluorescence, turbidity and pH will
be measured every 15 minutes during the test trials by two identical multi-parameter probes
(calibrated according to manufactures specification) placed, one each, into the control and test
tanks. A third hand-held instrument will be used to measure temperature, salinity, and dissolved
oxygen of water in each replicate sample (described above) as it is collected.
Initial inline samples (three replicates, 500 ml - 2 l each) of ballast water during the
filling of the control and test tanks will also be collected, filtered, and analyzed for the water
quality parameters of particulate organic carbon (POC), dissolved organic carbon (DOC), and
total suspended solids (TSS). See Appendices A, B and C for details.
Quantifying Viable Organism > 50 µm in size:
As described above, MERC uses five 1 m
3
mesocosms (a 5 m
3
integrated sample) and
GSI use three 1 m
3
mesocosms (a 3 m
3
integrated sample) to sample each time point and
treatment type (Table 2, page 27). Sampling occurs during initial uptake of water, just
downstream of the treatment systems during filling of the test tank, and upon discharge of
control and treated water (after 5 days). Immediately after filling, each mesocosm will be
drained through a 35 µm (50 µm diagonal dimension) plankton net to concentrate the
zooplankton for examination under a dissecting microscope. The proportion and total
concentration of live versus dead organisms will be determined using standard movement and
response to stimuli techniques and this live/dead analysis will take place within one hours of
collecting the individual samples. Depending on concentrations, quantification of zooplankton
in initial samples (upon ballasting) and control samples may require analysis of sub-samples and
extrapolation to the entire 1 m
3
. Zooplankton samples will then also be fixed with buffered, 10%
formalin in 125ml Nalgene bottles and shipped to the SERC for additional taxonomic
evaluations. Total counts and general taxonomic classification will be conducted under a
dissecting microscope at 25X, except for some taxa, which will be removed and identified using
a compound microscope. Larval forms of invertebrates will be identified to higher taxonomic
levels such as order (e.g., Decapoda) suborder (e.g., Balanomorpha) or class (e.g., Bivalvia).
Adults will be identified to species in most cases.
Quantifying Viable Organism 10 - 50 µm in size:
MERC - Two liters of unfiltered water for each mesocosm (a 10 l integrated sample) will
be collected immediately after filling, to determine concentrations of organisms in this size class
using four distinct methods (A – D below, Table 2 page 27). All samples will be held in amber
Nalgene bottles and transported on ice to laboratories where analyses occur within 3 hours of
collection. (A) One sub-sample from the initial 2 l will be fixed with standard Lugol’s solution,
and placed in a 250 ml amber Nalgene bottles to determine total cell abundances under an
inverted compound microscope using grid settlement columns and phase contrast lighting. (B) A